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FACULTY OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY DEPARTMENT OF PHYSIOLOGY AND PHARMACOLOGY UNIVERSIDAD NACIONAL AUTÓNOMA DE MÉXICO EVALUATION OF THE EFFECTIVENESS OF VIUSID-VET LIQUID* ON THE PRODUCTIVE PARAMETERS AND ALSO ITS IMMUNOMODULATORY ACTION ON BROILERS AT A COMMERCIAL POULTRY FARM Background Intensive broiler production nowadays not only means that the work involved in feeding, handling, biosafety and preventive medicine must be extremely efficient, but also the birds have to be kept healthy for as long as possible. Looking after these poultry populations and making sure that the maximum standards of health are upheld is a huge challenge and that is why growth enhancers and products that improve and preserve the birds’ overall general immune response (immunomodulators) are essential. With Viusid-Vet Liquid*, a commercial product from Catalysis distributed by Laboratorio Dermaceutical México, S. A. de C. V, the immune system of broilers and their productive expression can indeed be optimized. Viusid-Vet Liquid* is a food supplement, composed of antioxidants, vitamins, trace elements and an active substance from liquorice root extract (glycyrrhizinic acid) with potential antiviral properties. The molecular activation of its active substances stimulates their biological functions (antiviral and antioxidant effect) without modifying their molecular structure, which significantly builds up the organism’s defences. Objective Evaluate the effectiveness of Viusid-Vet Liquid* on the productive parameters and also how it works like an immunomodulator in broilers throughout the fattening period when it is added to their drinking water. Hypothesis Increase the humoral immune response when Viusid-Vet Liquid* is added to the drinking water and improve the productive parameters of the broilers. Material and methods Species: Gallus gallus male broilers, Ross x Ross strain, one day old and housed in two sheds: 21,415 birds in shed 2, treated with Viusid-Vet Liquid*, and 20,986 birds in the control group shed 3; 42,401 birds in total. Place: Josefina commercial poultry farm, located in Yautepec (Morelos, Mexico). The sheds are fitted out with an automatic feed system and cup nipple drinkers, and the birds are reared in semicontrolled conditions. 42,401 birds were used in total. Product: Viusid-Vet Liquid*. Commercial product from Catalysis, distributed by Laboratorio Dermaceutical México, S. A. de C. V. It is available in a metallic packet of 270 ml and it is added to the drinking water. Dose of 540 ml/1000 L of drinking water. Feed: Grain feed produced by the firm itself for different stages of the production cycle: Pre-starter, from 0-10 days; Starter, 11-8 days; Grower, 19-21 days; Finisher, from 28 to 45 days; and Withdrawal, from 45 days to market weight. Monitoring: 16 samples of serum and the lymphoid organs from 5 slaughtered birds were used to obtain histological sections for each group in the clinical trial. Evaluation time: From 1 day old to 49 days old. Groups: Birds in shed 2 received Viusid-Vet Liquid* that was added to the drinking water in a ratio of 540 ml / 1000 L continuously throughout the fattening period of 49 days. Birds in shed 3 were used as the control group. All the birds were given water and feed ad-libitum and they were vaccinated in accordance with the established vaccination calendars at the farm. • Data for the experimental design - Josefina farm - Number of birds in the clinical trial: 42,401 - Birds in the control group: 20,986 - Birds in the treatment group: 21,415 • Evaluated parameters at 35 days - % of mortality - Weekly weight - Cumulative weekly feed conversion Serological and histopathological tests - Haemagglutination-inhibition test for Newcastle’s disease (HI-ND) at 28 and 44 days old - Histopathological test on lymphoid organs (spleen, bursa of Fabricius and thymus) at 28 and 44 days old 16 samples of serum (16 s.) and 10 lymphoid organs (5 o.) were taken from birds of each group to carry out the tests. The following monitoring programme was established for the two groups, the control group and the group treated with Viusid-Vet Liquid: Days old Laboratory tests 28 days HI-ND (16 birds/group) 28 days Histopathological test: T, BF, S and L (5 birds/group) 44 days 44 days HI-ND (16 birds/group) Histopathological test: T, BF, S and L (5 birds/group) Haemagglutination-inhibition test for Newcastle’s disease T: Thymus, BF: Bursa of Fabricius, S: Spleen and L: Liver. Haemagglutination-inhibition test for Newcastle’s disease T: thymus, BF: Bursa of Fabricius, S: Spleen and L: Liver. Laboratory tests: Considering the immunomodulatory effect of Viusid-Vet Liquid*, the main objective was to see whether there was an increase in the number of cells, tissue and the weight of the lymphoid organs, which therefore involved calculating the weight of the lymphoid organs and the levels of antibodies against Newcastle’s disease (HI-ND). Vaccination calendar: Days old 1 day Type of vaccine MD+ IBD/ subcutaneous Combined live vaccine against Marek's disease and the infectious bursal disease. 6 days 8 days 10 days Newcastle / ocular IBD/ oral ND-IB/ ocular 10 days ND-AI/ subcutaneous 14 days IBD/ oral Live vaccine against Newcastle's disease, ocular administration Live vaccine against the infectious bursal disease Live vaccine against Newcastle's disease and Infectious bronchitis Inactivated vaccine against Newcastle's disease and Avian Influenza Live vaccine against the infectious bursal disease. Results It should be pointed out that there were respiratory problems during the cycle that became apparent with the number of deaths recorded at 29 days and from then after, for the birds in shed 2 that were being treated with Viusid-Vet Liquid. The infection had probably spread through all the sheds on the farm. The symptomatology, the mortality and the lesions observed all point to the fact that the respiratory disorders had been caused by an infection (infectious coryza) and by the subsequent more complex bacteria-related problems. The first serological markers are monitored in the initial stages of the clinical trial, which is when the respiratory symptomatology that persists right up until the end, is noticed. This infection is thought to have altered the productive parameters, in particular the cumulative and weekly mortality parameters. Productive parameters recorded at Josefina farm for the broilers that are continuously treated with Viusid-Vet Liquid* 540 ml/1000 L of drinking water, until they are 49 days old. Weight, mortality and feed conversion are as follows: Week 1 2 3 4 5 6 7 VIUSID Weight 0.135 0.356 0.672 1.180 1.615 2.164 2.750 Control Weight 0.132 0.320 0.675 1.190 1.640 2.130 2.520 % 2% 11% 0% -1% -2% 2% 9% VIUSID %C. Mort. 1.251 2.260 3.054 3.960 5.870 9.647 15.970 Control % C. Mort. 1.010 2.363 3.402 4.336 6.428 12.785 19.718 % 24% -4% -10% -9% -9% -25% -19% VIUSID F. Conv. 1.680 1.744 1.486 1.425 1.772 1.944 2.079 Control F.Conv. 1.513 1.768 1.348 1.325 1.679 1.992 2.260 % 11% -1% 10% 8% 4% -2% -8% With regard to the average weights, at the end of the clinical trial, the group treated with Viusid-Vet Liquid* weighed 9% more, namely 230 grams, than the birds from the control group. This is a considerable difference in statistical terms. The average cumulative and weekly mortality was always lower for the group treated with Viusid-Vet Liquid* from the second week up to the end of the clinical trial, as mortality dropped 19% (3.748 real points) compared to that of the birds in the control group (P<0.05). The average feed conversion at the end of the clinical trial for the Viusid-Vet Liquid* treatment group was 8% better, which is equivalent to 190 g of feed per kilo of weight, compared to that of the birds in the control group. This is also a considerable difference in statistical terms. Histopathological results Weight of organs The average weight of the organs when the birds were 28 days old was better for the birds treated with Viusid-Vet liquid* except for the thymus weight. Sample Average Spleen (g) 0.99 Thymus (g) 0.69 Control group Bursa of Fabricius (g) 0.66 Liver (g) 14.99 Sample Average Spleen (g) 1.18 Viusid-Vet Liquid* group Thymus (g) Bursa of Fabricius (g) 0.57 0.78 Liver (g) 20.18 Positive differences were observed in the group treated with Viusid-Vet Liquid*; due to the existence of lymphoid hyperplasia and the fact that there were fewer lesions in their organs compared to those from the control group. The histopathological results of the lymphoid organs corroborate the immunostimulatory effect of Viusid-Vet Liquid*. Spleen Moderate lymphoid hyperplasia Abundant lymphoid hyperplasia No significant changes Small areas of dispersed focal necrosis Slight lymphoid hyperplasia Control group 28 days VIUSID 28 days 0% 100% 0% 0% 100% 0% Control group 44 days VIUSID 44 days 60% 60% 20% 0 20% 40% Spleen % of positive samples Moderate lymphoid hyperplasia Abundant lymphoid hyperplasia No significant changes Control group 28 days old VIUSID 28 days old The moderate lymphoid tissue development of the spleen in birds from the Viusid Vet Liquid* treatment group was better than that of the control group in 100% of the samples. Spleen % of positive samples Small areas of dispersed focal necrosis Slight lymphoid hyperplasia No significant changes Control group 44 days old VIUSID 44 days old The lymphoid tissue development of the spleen in birds from the Viusid Vet Liquid* treatment group was better than that of the control group, in which 20% of the birds had tissue affected by small areas of focal necrosis. Control group 28 days VIUSID 28 days Control group 44 days VIUSID 44 days Liver Slight swelling Moderate swelling No significant changes Slight focal lymphoid hyperplasia Severe swelling 60% 0% 40% 40% 40% 20% 20% 0% 20% 60% 0% 40% 60% 0% ***At 28 days there were no significant changes in the aforesaid liver lesions of the birds from either group involved in this clinical trial. Liver % of positive samples Slight focal lymphoid hyperplasia Slight swelling Severe swelling No significant changes VIUSID 44 days old Control group 44 days old N.B.: The growth of the lymphoid tissue and the reduction of lesions in the liver were better in the group treated with Viusid-Vet Liquid* than the control group. ***There were no significant differences in the thymus lesions described in the histological study when the birds were 28 and 44 days old for either the control group or the group treated with ViusidVet Liquid*. Bursa of Fabricius Moderate subacute bursitis Severe bursitis Severe chronic bursitis Moderate chronic bursitis Chronic slight with areas of cortical regeneration Moderate cortical hyperplasia Moderate follicular atrophy with areas of cortical regeneration Control group 28 days 0% 0% 100% VIUSID 28 days 60% 20% 20% Control group 44 days VIUSID 44 days 20% 0% 80% 0% 20% 0% 0% 20% 0% 60% 20% 0% Bursa of Fabricius % of positive samples Moderate subacute bursitis Severe bursitis Severe chronic bursitis Control group 28 days VIUSID 28 days When the birds from the group treated with Viusid-Vet Liquid* were 28 days old they had fewer chronic inflammatory lesions, 80% of the birds had no lesions at all. This means that they had more protection than the birds from the control group, in which 100% of the birds had chronic inflammatory lesions. The lymphoid tissue development was 60% better in the birds from the group that was administered Viusid-Vet Liquid* when they were 44 days old compared to that of the birds in the control group. The cortical regeneration was 20% better in the birds from the group treated with Viusid-Vet Liquid* than that of the birds in the control group. Serological results Inhibition test for Newcastle’s disease (HI-ND) at 28 days old Dilution 2 4 16 32 64 128 256 512 Control group 28 days 1 1 0 2 2 6 3 1 HI for Newcastle VIUSID 28 days Control group 44 days 0 0 0 0 0 1 5 0 5 3 4 6 1 4 2 0 VIUSID 44 days 0 0 0 0 0 7 5 2 28 days Control group 28 days VIUSID group N.B.: The protective antibody titre levels of two of the serum samples taken from birds in the control group were lower than the minimum, whilst those in the birds from the group treated Viusid-Vet Liquid* were, on average, higher and uniform. HI for Newcastle’s disease Nº of positives Dilution Control 44 days VIUSID 44 days The serological results (haemagglutination-inhibition test) for Newcastle’s disease (ND), using 10 haemagglutination units and expressed in the base two logarithmic average, were better for the birds from the treatment group administered Viusid-Vet Liquid* when they were 6.2 weeks old: Age (days) 28 44 Control group 28 days VIUSID group 28 days Control group 44 days VIUSID group 44 days Control Group Log 2 6.25 6.86 Viusid Group Log 2 6.41 7.64 The infection that was observed in both groups could have affected the antibody titres against ND. The impact that this might have had at 21 days old, which produced a log of 0.16 and at 44 days old a log of 0.78, is unknown, although the antibody titre levels of the birds treated with Viusid-Vet Liquid* were always higher than those of the birds in the control group. Conclusions 1. The resulting productive parameters of the birds treated with Viusid-Vet Liquid* proved its immunomodulatory effect on the lymphoid organs. 2. Viusid-Vet Liquid* significantly improved the productive parameters concerning the feed conversion, weight and mortality rates of the broilers in the treatment group, despite the fact that they had an infection. 3. Viusid-Vet Liquid* produced more protective, homogenous and uniform antibody titres against Newcastle’s disease (HI-ND) in the birds from the treatment group when they were 28 and 44 days old compared to those in the control group. 4. Viusid-Vet Liquid* favoured the weight and the cell levels in the primary lymphoid tissues, namely, the spleen, bursa of Fabricius and liver. Adding Viusid-Vet Liquid* to the birds' drinking water does not alter their homeostasis or their symptomatology in any way after the product has been used. It does however improve the productive expression in strains of high yield broilers. *Registered trade mark Viusid-Vet Liquid* by Catalysis (Spain). Distributed by Dermaceutical México, S. A. de C. V. KINDS REGARDS LUIS OCAMPO CAMBEROS Doctor of Veterinary Science and Animal Husbandry FULL TIME PROFESSOR DEPARTMENT OF PHARMACOLOGY FACULTY OF VETERINARY SCIENCE AND ANIMAL HUSBANDRY (FMVZ) UNIVERSIDAD NACIONAL AUTONOMA DE MEXICO (UNAM) Bibliography